Microenvironment of Endosomal Aqueous Phase Investigated by the Mobility of Microparticles Using Fluorescence Correlation Spectroscopy

Yoshida, N.; Kinjo, Masataka; Tamura, Mamoru

doi:10.1006/bbrc.2000.4115

Cited by 39 publications

(29 citation statements)

References 32 publications

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“…When the confocal volume is deep in the solution (z = 10 μm), the ACF decays with a single decay characteristic and is well described by equation (2). When the confocal volume contains the water-surface interface, the ACF decay has a slow time contribution.…”

Section: Resultsmentioning

confidence: 99%

“…Fluorescence correlation spectroscopy (FCS) has become the method of choice for investigating various molecular properties such as diffusion, [1][2][3][4][5][6][7] the conformational dynamics of proteins, [8][9][10] and binding and reaction kinetics [10][11][12][13] at the single-molecule level due to the development of excellent laser sources with high beam quality and stability, sophisticated detectors with low noise and high sensitivity, development of objectives with long working distances and high numerical apertures (NAs), and high-quality optics. 14,15 In FCS, signals are recorded as fluctuations in the fluorescence signal from a molecule passing through a detection volume, which is on the order of femtoliters.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

Chowdhury¹,

Manho²

2011

Bulletin of the Korean Chemical Society

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Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

Chowdhury¹,

Manho²

2011

Bulletin of the Korean Chemical Society

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“…Fluorescence autocorrelation curves G () were fitted to a singlecomponent diffusion model (Yoshida et al, 2001;Jin et al, 2007) as follows: Therefore, the long-term diffusion coefficient for the specific time period could be directly obtained from this plot (Chung et al, 2010).…”

Section: Methodsmentioning

confidence: 99%

Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging

Nishimura¹,

Ritchie²,

Kasai³

et al. 2013

J Gen Physiol

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“…However, viscosity differs among the individual cellular organelles. While the viscosity in endosomes, [47] for example, is only slightly greater than the viscosity of the cytoplasm, the viscosity of the nucleus seems significantly increased. [48,49] In addition, the viscosity of the organelles and the cytoplasm can also change during different states of the cell, for example, different phases of the cell cycle.…”

Section: ) Labelingmentioning

confidence: 95%

In‐Cell NMR Spectroscopy

2005

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Microenvironment of Endosomal Aqueous Phase Investigated by the Mobility of Microparticles Using Fluorescence Correlation Spectroscopy

Cited by 39 publications

References 32 publications

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging

In‐Cell NMR Spectroscopy

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